The innate antiviral immune system of the cat: molecular tools for the measurement of its state of activation

The innate immune system plays a central role in host defence against viruses. While many studies portray mechanisms in early antiviral immune responses of humans and mice, much remains to be discovered about these mechanisms in the cat. With the objective of shedding light on early host-virus interactions in felids, we have developed 12 real-time TaqMan(?) qPCR systems for feline genes relevant to innate responses to viral infection, including those encoding for various IFNα and IFNω subtypes, IFNβ, intracellular antiviral factor Mx, NK cell stimulator IL-15 and effectors perforin and granzyme B, as well as Toll-like receptors (TLRs) 3 and 8. Using these newly developed assays and others previously described, we measured the relative expression of selected markers at early time points after viral infection in vitro and in vivo. Feline embryonic fibroblasts (FEA) inoculated with feline leukemia virus (FeLV) indicated peak levels of IFNα, IFNβ and Mx expression already 6h after infection. In contrast, Crandell-Rees feline kidney (CrFK) cells inoculated with feline herpes virus (FHV) responded to infection with high levels of IFNα and IFNβ only after 24h, and no induction of Mx could be detected. In feline PBMCs challenged in vitro with feline immunodeficiency virus (FIV), maximal expression levels of IFNα, β and ω subtype genes as well as IL-15 and TLRs 3, 7 and 8 were measured between 12 and 24h after infection, whereas expression levels of proinflammatory cytokine gene IL-6 were consistently downregulated until 48h post inoculation. A marginal upregulation of granzyme B was also observed within 3h after infection. In an in vivo experiment, cats challenged with FIV exhibited a 2.4-fold increase in IFNα expression in blood 1 week post infection. We furthermore demonstrate the possibility of stimulating feline immune cells in vitro with various immune response modifiers (IRMs) already known for their immunostimulatory properties in mice and humans, namely Poly IC, Resiquimod (R-848) and dSLIM?, a synthetic oligonucleotide containing several unmethylated CpG motifs. Stimulation of feline PBMCs with dSLIM? and R-848 effectively enhanced expression of IFNα within 12h by factors of 6 and 12, respectively, and Poly IC induced an increase in Mx mRNA expression of 28-fold. Altogether, we describe new molecular tools and their successful use for the characterization of innate immune responses against viruses in the cat and provide evidence that feline cells can be stimulated by synthetic molecules to enhance their antiviral defence mechanisms.     

First morphological characterization of 'Candidatus Mycoplasma turicensis' using electron microscopy

At least three haemotropic mycoplasmas have been recognized in cats: Mycoplasma haemofelis (Mhf), 'Candidatus Mycoplasma haemominutum' (CMhm) and 'Candidatus M. turicensis' (CMt). The latter was originally identified in a Swiss pet cat with haemolytic anaemia and shown to be prevalent in domestic cats and wild felids worldwide using molecular methods. So far, there has been no confirmatory morphological evidence of the existence of CMt presumably due to low blood loads during infection while CMhm has only been characterized by light microscopy with discrepant results. This study aimed to provide for the first time electron microscopic characteristics of CMt and CMhm and to compare them to Mhf. Blood samples from cats experimentally infected with CMt, CMhm and Mhf were used to determine copy numbers in blood by real-time PCR and for transmission and scanning electron microscopy. High resolution scanning electron microscopy revealed CMt and CMhm to be discoid-shaped organisms of 0.3 μm in diameter attached to red blood cells (RBCs). In transmission electron microscopy of CMt, an oval organism of about 0.25 μm with several intracellular electron dense structures was identified close to the surface of a RBC. CMhm and CMt exhibited similar morphology to Mhf but had a smaller diameter. This is the first study to provide morphological evidence of CMt thereby confirming its status as a distinct haemoplasma species, and to present electron microscopic features of CMhm.     

Evaluating the protective capacity of two-post ROPS for a seat-belted occupant during a farm tractor overturn

This study evaluated the effectiveness of a commercial rollover protective structure (ROPS) and size-extended ROPS in protecting a 95th percentile male operator during tractor overturns. Six rear upset tests (commercial ROPS) and ten side upset tests (commercial and size-extended ROPS) were conducted. A 95th percentile instrumented male manikin was used in all tests. Head injury criterion (HIC15), 80 g limit on resultant head acceleration, neck injury criterion (Nij), and peak axial force (extension-compression) were employed to evaluate injury potential. In all rear tests, the manikin's head impact with the ground was within the tolerance limits for head/neck injuries. Based on limited trials in the side tests, the study observed a small to moderate chance of neck injuries under the commercial and size-extended ROPS conditions; the injury risk was not statistically significant between these two test conditions. This study identified a risk of non-fatal injuries for large-size operators in side overturns, although the prevention effectiveness of commercial versus size-extended ROPS cannot be determined without further testing. These findings may have implications for future ROPS designs.     

Guideline for the description of soils in the Berlin metropolitan area: an extension for surveying and mapping anthropogenic and natural soils in urban environments within the German soil classification system

Journal of Soils and Sediments - In urban areas, humans shape the surface, (re-)deposit natural or technogenic material, and thus become the dominant soil formation factor. The 2015 edition of the...     

Die Mitzweisung der Magnetnadel

Ohne Zusammenfassung     

Effect of fertilizers and irrigation on multi-configuration electromagnetic induction measurements

Electromagnetic induction (EMI) data are often used to investigate spatial and temporal patterns of soil texture, soil water content and soil salinity. We hypothesized that the EMI methodology might thus also offer potential to detect agricultural legacy effects originating from fertilizer application and irrigation of different fields. Therefore, we performed EMI measurements on two long-term field experiments (LTFE) at Thyrow near Berlin (Germany) that differed in agricultural management with regard to long-term irrigation in combination with mineral (NPK and lime) and organic amendments (straw and farmyard manure). Two different rigid-boom multi-coil EMI instruments were used to measure simultaneously the apparent electrical conductivity (ECa) over nine different depth ranges to study the entire soil profile from topsoil to deep subsoil. Additionally, soil samples were taken from the different treatments to ground-truth the measurements and disentangle the nutrient application or irrigation effects from natural soil heterogeneity. The soil samples indicated a rather homogenous soil and the correlation between soil parameters or states were not significant. However, the treatments showed significant differences in measured ECa values. In general, ECa values were largest on regularly irrigated as well as on mineral and organic fertilized plots, with regular irrigation exhibiting the largest impact on EMI records even though the last application was months before the EMI measurement. Overall, this study reveals that EMI data can support the classical in situ assessment of agricultural management effects within LTFE, while offering new potentials in detecting and understanding legacy effects of agricultural management on spatial soil properties at farm level.